Writing a Custom Affinity Method Using Scouting for the AKTA Avant Chromatography System.

In this video we write a Unicorn Method from scratch, without using any of the preset phases. The method includes Scouting and is meant to be run on an AKTA Avant Chromatography system.
It's a long video so get some popcorn and get ready to nerd out over chromatography. If you ever actually watch it leave a comment, I'd be grateful.
How to setup and some example results can be found at: • Setting Up a Custom Af...
One of these days I'll learn how to spell equilibration properly, but I don't think it matters for the purpose of this video to have it absolutely correct.
Here is the text of the method:
UNICORN 7.6.0
Method: MabSelect method
Text instructions
Main method:
0.00 Base: CV, Vc=7.854 {ml}, (Mabselect tricorn column)#Column type
0.00 Phase: Method Settings
0.00 Base: SameAsMain
0.00 Alarm pre column pressure: Enabled, (0.75)#Pre column pressure limit {MPa}, 0.00 {MPa}
0.00 Alarm delta column pressure: Enabled, 0.75 {MPa}, 0.00 {MPa}
0.00 Wavelength: (280)#UV1 {nm}, Off {nm}, Off {nm}
0.00 Noise reduction UV: (0.2)#UV averaging time {sec}
0.00 Injection valve: Manual load
0.00 Outlet valve: Out-Waste
0.00 pH valve: (In-line)#pH cell, (In-line)#Flow restrictor
0.00 Alarm air sensors: (Enabled)#Air sensor alarm on inlet valve A,
(Enabled)#Air sensor alarm on inlet valve B, (Disabled)#Air sensor alarm
on sample inlet valve
0.00 Inlet A: (A1)#Wash Buffer
0.00 Inlet B: (B1)#Elution Inlet
0.00 Pump wash: (Off)#Wash A, (Off)#Wash B, Off, (Buffer)#Wash Sample
0.00 Block: Mehtod Flowrate
0.00 Base: SameAsMain
0.00 System flow: (3.600)#Flow rate {ml/min}, (Pre column pressure)
#Pressure control
0.00 End_Block
0.00 End_Block
0.00 Phase: Column Equilibration
0.00 Base: SameAsMain
0.00 Block: Delay A
0.00 Base: Volume, Any
10.00 End_Block
0.00 Column position: (Position 1)#Column position, Down flow
(1.00)#Length of Equilibration End_Block
0.00 Phase: Load Cell Harvest
0.00 Base: Volume, Any
0.00 System flow: 0.000 {ml/min}, Off
0.00 Block: Delay B
0.00 Base: Time, Any
0.05 End_Block
0.00 Injection valve: Direct inject
0.00 Sample inlet: (S1)#Sample Inlet position
0.00 Injection mark
0.00 Sample flow: (3.600)#Sample flowrate {ml/min}, Off
0.00 Outlet valve: (Out-Waste)#Cell Harvest Waste
0.00 Hold until: Air sensor Sample inlet valve, Equals, Air, (500.00)
#Cell Harvest Load volume {base}
0.02 Block: Reset system
0.00 Base: SameAsMain
0.00 Sample flow: 0.000 {ml/min}, Off
0.00 Block: Delay C
0.00 Base: Time, Any
0.05 End_Block
0.00 Injection valve: Manual load
0.00 Block: Mehtod Flowrate
0.00 Base: SameAsMain
0.00 System flow: (3.600)#Flow rate {ml/min}, (Pre column
pressure)#Pressure control
0.00 End_Block
0.00 End_Block
0.02 End_Block
0.00 Phase: Wash Column
0.00 Base: SameAsMain
1.00 Outlet valve: Out-Waste
4.00 End_Block
0.00 Phase: Elution
0.00 Base: SameAsMain
0.00 Gradient: 100.0 {%B}, 0.00 {base}
0.00 Block: Delay D
0.00 Base: SameAsMain
0.10 End_Block
0.00 Auto zero UV
0.00 Block: Delay E
0.00 Base: Time, Any
0.05 End_Block
0.00 Peak fractionation parameters: Level, 0.150 {min}, (100.000)#Start
Peak Frac {mAU}, 100.000 {mAU/min}, (75.000)#End Peak Frac {mAU}, 75.000
{mAU/min}
0.00 Peak fractionation: Volume, 15 ml tubes, 12.0 {ml}, Next tube
3.00 End_Block
0.00 Phase: Column Cleaning
0.00 Base: SameAsMain
0.00 Gradient: 0.0 {%B}, 0.00 {base}
0.00 Stop peak fractionation
0.00 Block: (CIP)#Skip or CIP
0.00 Base: Time, Any
0.00 System flow: 2.500 {ml/min}, Off
0.00 Inlet A: (A2)#CIP solution
30.00 End_Block0.00 End_Block
0.00 Phase: Column Reequilibration
0.00 Base: SameAsMain
0.00 Inlet A: A1
1.00 Block: Mehtod Flowrate
0.00 Base: SameAsMain
0.00 System flow: (3.600)#Flow rate {ml/min}, (Pre column pressure)#Pressure control
0.00 End_Block
(5.00)#Length of reequlibration End_Block
(Unused)
0.00 Block: Skip
0.00 Base: SameAsMain
0.00 End_Block
A note on delay blocks (if you've come this far): I tested this method and it doesn't look like delay blocks B to E are necessary. These used to be necessary (Unicorn version 3 & 5 needed them), I think the authors of the Unicorn software added some features in either 6 or & 7 that make it less necessary. I decided to leave them in however since I still believe it will increase fidelity and demonstrates problem solving. Take it for what you will.
Reach me at my Linkedin page / chad-blamey-ph-d-649a336
Music used under Creative Commons license:
Music Credit: LAKEY INSPIRED
Track Names: "Better Days" "Chill Days" "Warm nights" "Blue Boi"
Music By: LAKEY INSPIRED @ / lakeyinspired
Also "Daisy" by Chillpeach • Video
Thanks for letting me use this music!